Y-27632 enables long-term expansion of mouse submandibular gland epithelial cells via inactivation of TGF-ß1/CTGF/p38 and ROCK2/JNK signaling pathway.

OBJECTIVES: This study aimed to investigate the effects of Y-27632 on the long-term maintainence of mouse submandibular epithelial cells (SG-Epis) in vitro and to elucidate the underlying mechanisms. METHODS: The role of the Rho-associated kinase (ROCK) inhibitor Y-27632 in maintaining SG-Epis and its underlying mechanisms were evaluated by examining the in vitro expansion of mouse SG-Epis. Changes in key cellular characteristics, such as proliferation, long-term expansion, and mRNA and protein expression, were assessed in the presence or absence of Y-27632. RESULTS: Treatment with Y-27632 significantly enhanced the proliferative potential of SG-Epis, preserving Krt8 and Krt14 expression over 17 passages. In the absence of Y-27632, SG-Epis lost their epithelial morphology. However, Y-27632 treatment maintained the epithelial morphology and downregulated mRNA levels of Tgf-ß1, Ctgf, and Rock2. Treatment with TGF-ß1 indicated that TGF-ß/CTGF/p38 signaling is responsible for the maintenance of SG-Epis, while RNA interference studies revealed that ROCK2/c-Jun N-terminal kinase (JNK) signaling is also crucial for SG-Epis proliferation and maintenance. CONCLUSIONS: The TGF-ß1/CTGF/p38 and ROCK2/JNK signaling pathways are responsible for SG-Epis proliferation, and Y-27632 treatment effectively inactivates these pathways, enabling long-term in vitro maintenance of SG-Epis. The culture method utilizing Y-27632 provides an effective approach for the in vitro expansion of SG-Epis.

Ultra-Wideband Electromagnetic Composite Absorber Based on Pixelated Metasurface with Optimization Algorithm.

An ultra-wideband electromagnetic (EM) absorber is proposed. The proposed absorber consists of two thin metasurfaces, four dielectric layers, a glass fiber reinforced polymer (GFRP), and a carbon fiber reinforced polymer (CFRP) which works as a conductive reflector. The thin metasurfaces are accomplished with 1-bit pixelated patterns and optimized by a genetic algorithm. Composite materials of GFRP and CFRP are incorporated to improve the durability of the proposed absorber. From the full-wave simulation, more than 90% absorption rate bandwidth is computed from 2.2 to 18 GHz such that the fractional bandwidth is about 156% for the incidence angles from 0° to 30°. Absorptivity is measured using the Naval Research Laboratory (NRL) arch method in an EM anechoic environment. It was shown that the measured results correlated with the simulated results. In addition, the proposed absorber underwent high temperature and humidity tests under military environment test conditions in order to investigate its durability.

Fabrication of junction-free Cu nanowire networks via Ru-catalyzed electroless deposition and their application to transparent conducting electrodes.

Over the past few years, metal nanowire networks have attracted attention as an alternative to transparent conducting oxide materials such as indium tin oxide for transparent conducting electrode applications. Recently, electrodeposition of metal on nanoscale template is widely used for formation of metal network. In the present work, junctionless Cu nanowire networks were simply fabricated on a substrate by forming a nanostructured Ru with 80 nm width as a seed layer, followed by direct electroless deposition of Cu. By controlling the density of Ru nanowires or the electroless deposition time, we readily achieve desired transmittance and sheet resistance values ranging from ∼1 kΩ sq-1at 99% to 9 Ω sq-1at 89%. After being transferred to flexible substrates, the nanowire networks exhibited no obvious increase in resistance during 8000 cycles of a bending test to a radius of 2.5 mm. The durability was verified by evaluation of its heating performance. The maximum temperature was greater than 180 °C at 3 V and remained constant after three repeated cycles and for 10 min. Transmission electron microscopy and x-ray diffraction studies revealed that the adhesion between the electrolessly deposited Cu and the seed Ru nanowires strongly influenced the durability of the core-shell structured nanowire-based heaters.

A Rho Kinase (ROCK) Inhibitor, Y-27632, Inhibits the Dissociation-Induced Cell Death of Salivary Gland Stem Cells.

Salivary gland stem cells (SGSCs) are potential cell sources for the treatment of salivary gland diseases. The control of cell survival is an essential factor for applying stem cells to regenerative medicine or stem cell-based research. The purpose of this study was to investigate the effects of the ROCK inhibitor Y-27632 on the survival of SGSCs and its underlying mechanisms. SGSCs were isolated from mouse submandibular glands and cultured in suspension. Treatment with Y-27632 restored the viability of SGSCs that was significantly decreased during isolation and the subsequent culture. Y-27632 upregulated the expression of anti-apoptotic protein BCL-2 in SGSCs and, in the apoptosis assay, significantly reduced apoptotic and necrotic cell populations. Matrigel was used to mimic the extracellular environment of an intact salivary gland. The expression of genes regulating apoptosis and the ROCK signaling pathway was significantly reduced when SGSCs were embedded in Matrigel. SGSCs cultured in Matrigel and treated with Y-27632 showed no difference in the total numbers of spheroids and expression levels of apoptosis-regulating genes. Matrigel-embedded SGSCs treated with Y-27632 increased the number of spheroids with budding structures and the expression of acinar cell-specific marker AQP5. We demonstrate the protective effects of Y-27632 against dissociation-induced apoptosis of SGSCs during their culture in vitro.

Slim-panel holographic video display.

Since its discovery almost 70 years ago, the hologram has been considered to reproduce the most realistic three dimensional images without visual side effects. Holographic video has been extensively researched for commercialization, since Benton et al. at MIT Media Lab developed the first holographic video systems in 1990. However, commercially available holographic video displays have not been introduced yet for several reasons: narrow viewing angle, bulky optics and heavy computing power. Here we present an interactive slim-panel holographic video display using a steering-backlight unit and a holographic video processor to solve the above issues. The steering-backlight unit enables to expand the viewing angle by 30 times and its diffractive waveguide architecture makes a slim display form-factor. The holographic video processor computes high quality holograms in real-time on a single-chip. We suggest that the slim-panel holographic display can provide realistic three-dimensional video in office and household environments.

Newly synthesized peptide, Ara-27, exhibits significant improvement in cell-penetrating ability compared to conventional peptides.

Cell-penetrating peptides (CPPs) are short amino acid sequences known to act as a vehicle for enhancing the intracellular translocating efficiency of extracellular molecules. Although many groups have attempted to develop peptides with high cell-penetrating efficiencies, very few have demonstrated efficient cellular uptake of CPPs at low concentrations. Here, we describe a newly synthesized peptide derived from Arabidopsis, Ara-27, which exhibits significant improvement in cell-penetrating efficiency compared to existing CPPs. The cell-penetrating efficiency of Ara-27 was compared with the commonly used Tat-protein transduction domain (Tat-PTD) and membrane translocating sequence (MTS) in human dermal fibroblast (HDF) and human dental pulp stem cells (hDPSC). Cell-penetrating efficiency of fluorescein isothiocyanate (FITC)-labeled CPPs were assessed by flow cytometry and visualized by confocal microscopy. Flow cytometric analysis revealed >99% cell-penetrating efficiency for 2 µM Ara-27 in both HDF and hDPSC. In contrast, 2 µM Tat-PTD and MTS showed <10% cell-penetrating efficiency in both cells. In support, relative fluorescence intensities of FITC-labeled Ara-27 were around 8 to 22 times higher than those of Tat-PTD and MTS in both cells. Confocal analysis revealed internalization of 0.2 and 2 µM Ara-27 in both human cells, which was not observed for Tat-PTD and MTS at either concentration. In conclusion, this study describes a novel CPP, Ara-27, which exhibit significant improvement in intracellular uptake compared to conventional CPPs, without affecting cell viability. Thus, development of Ara-27 based peptides may lead to improved delivery of functional cargo such as small molecules, siRNA, and drugs for in vivo studies.

Enterococcus faecium L-15 Extract Enhances the Self-Renewal and Proliferation of Mouse Skin-Derived Precursor Cells.

Lactic acid bacteria (LAB) in the gastrointestinal tract have beneficial health effects. LAB activate the proliferation of intestinal stem cells and speed the recovery of damaged intestinal cells, but little is known about effect of LAB on other adult stem cells. In this study, a cell-free extract of Enterococcus faecium L-15 (L15) was exposed to mouse skin-derived precursor cells (SKPs), and the changes in characteristics associated with proliferation and self-renewal capacity were investigated. L15 increased the size of the spheres and the proliferation rate of SKPs. Cell cycle analysis revealed that cells in the S-phase increased after treatment with L15. In the L15-treated group, the total number of spheres significantly increased. The expression level of pluripotency marker genes also increased, while the mesenchymal lineage-related differentiation marker genes significantly decreased in the L15-treated group. The PI3K/Akt signaling pathway was activated by L15 in SKPs. These results indicate that L15 enhances proliferation and self-renewal of SKPs and may be used as a supplement for stem cell maintenance or application of stem cell therapy. This is the first report to investigate the functional effects of E. faecium on the proliferation and self-renewal capacity of SKPs.

A Low-Profile High-Gain and Wideband Log-Periodic Meandered Dipole Array Antenna with a Cascaded Multi-Section Artificial Magnetic Conductor Structure.

This paper presents a low-profile log-periodic meandered dipole array (LPMDA) antenna with wideband and high gain characteristics. The antenna consists of 14 dipole elements. For compactness, a meander line structure is applied to each dipole element to reduce its physical length. As a result, a compact and wideband LPMDA antenna is realized, exhibiting a wide impedance bandwidth of 1.04-5.22 GHz (ratio bandwidth of 5.02:1) for | S 11| < -10 dB. To enhance the antenna gain performance while maintaining the wideband behavior, the LPMDA antenna is integrated with a new design of an artificial magnetic conductor (AMC) structure. The designed AMC is realized by combining three AMC structures of different sizes to form a cascaded multi-section AMC structure, of which its overall operating bandwidth can continuously cover the entire impedance bandwidth of the LPMDA antenna. The proposed AMC-backed LPMDA antenna is experimentally verified and its measured -10 dB reflection bandwidth is found to be in the range of 0.84-5.15 GHz (6.13:1). At the main beam direction within the operating frequency bandwidth, the gain of the proposed AMC-backed LPMDA antenna ranges from 7.15-11.43 dBi, which is approximately 4 dBi higher than that of an LPMDA antenna without an AMC. Moreover, the proposed antenna has a low profile of only 0.138 λ L. ( λ L is the free-space wavelength at the lowest operating frequency).

Clinical experience with multiplex ligation-dependent probe amplification for microdeletion syndromes in prenatal diagnosis: 7522 pregnant Korean women.

BACKGROUND: Conventional cytogenetic analysis using G-band karyotyping has been the method of choice for prenatal diagnosis, accurately detecting chromosomal abnormalities larger than 5 Mb. However, the method is inefficient for detecting the submicroscopic deletions and duplications that are associated with malformations and mental retardation. This study evaluated the results of the multiplex ligation-dependent probe amplification (MLPA) P245 assay used for prenatal diagnosis in cases with unusual ultrasonographic findings or specifically where parents wanted to be tested. The objective was to compare the results from MLPA with those from conventional cytogenetic testing in order to determine their concordance and the additional diagnostic yield of MLPA over G-band karyotyping. RESULTS: Of the 7522 prenatal cases analyzed, 124 were found to have genomic imbalances (1.6%). Of those 124 cases, 41 had gene loss (33.6%), and 83 had gene gain (66.4%). Most of the cases with genomic imbalances (64.5%) showed no abnormal karyotype. In particular, all cases with a 4p16.3 deletion (Wolf-Hirschhorn syndrome) showed an abnormal karyotype, whereas all of those with a 22q11-13 deletion showed a normal karyotype. In most of the cases with pathogenic deletions, the indication for invasive prenatal testing was an increase in the nuchal translucency (NT) alone (51.2%). Other indications observed in the remaining cases were abnormal serum screening markers (14.6%), other ultrasonographic findings (9.8%), pregnancy through in vitro fertilization and fertility assistance (9.8%), and advanced maternal age(2.4%). CONCLUSIONS: These results show that for fetuses with an enlarged NT or abnormal ultrasonographic findings and normal conventional karyotype, additional genetic investigation like molecular testing would be for identifying the microscopic genomic aberrations (microdeletions, microduplications) responsible for syndromic associations including structural anomalies and mental retardation.

Enterococcus faecium L-15 Cell-Free Extract Improves the Chondrogenic Differentiation of Human Dental Pulp Stem Cells.

Hyaline cartilage is a tissue of very low regenerative capacity because of its histology and limited nutrient supply. Cell-based therapies have been spotlighted in the regeneration of damaged cartilage. Dental pulp stem cells (DPSCs) are multipotent and are easily accessible for therapeutic purposes. In human gastrointestinal tracts, Enterococcus faecium is a naturally occurring commensal species of lactic acid bacteria. In this work, the human DPSCs were differentiated into chondrocytes using a chondrogenic differentiation medium with or without L-15 extract. We observed that chondrogenic differentiation improved in an E. faecium L-15 extract (L-15)-treated DPSC group via evaluation of chondrogenic-marker mRNA expression levels. In particular, we found that L-15 treatment promoted early-stage DPSC differentiation. Cells treated with L-15 were inhibited at later stages and were less likely to transform into hypertrophic chondrocytes. In L-15-treated groups, the total amount of cartilage extracellular matrix increased during the differentiation process. These results suggest that L-15 promotes chondrogenic differentiation, and that L-15 may be used for cartilage repair or cartilage health supplements. To our knowledge, this is the first report demonstrating the beneficial effect of L-15 treatment on chondrogenic differentiation.

Sonic hedgehog signalling regulates the self-renewal and proliferation of skin-derived precursor cells in mice.

OBJECTIVES: The sonic hedgehog (Shh) signalling pathway has an important role in the maintenance of various stem cells and organogenesis during development. However, the effect of Shh in skin-derived precursors (SKPs), which have the capacity for multipotency and self-renewal, is not yet clear. The present study investigated the effects of the Shh signalling pathway on the proliferation and self-renewal of murine SKPs (mSKPs). METHODS: The Shh signalling pathway was activated by treatment with purmorphamine (Shh agonist) or recombinant Shh in mSKPs. Cyclopamine (Shh antagonist) or GANT-61 (Gli inhibitor) was used to inhibit the pathway. Western blot, qPCR, and immunofluorescence were used to analyse the expression of genes related to self-renewal, stemness, epithelial-mesenchymal transition (EMT) and the Shh signalling pathway. In addition, cell proliferation and apoptosis were examined. RESULTS: Inhibiting the Shh signalling pathway reduced mSKP proliferation and sphere formation, but increased apoptosis. Activating this signalling pathway produced opposite results. The Shh signalling pathway also controlled the EMT phenotype in mSKPs. Moreover, purmorphamine recovered the self-renewal and proliferation of aged mSKPs. CONCLUSION: Our results suggest that the Shh signalling pathway has an important role in the proliferation, self-renewal and apoptosis of mSKPs. These findings also provide a better understanding of the cellular mechanisms underlying SKP self-renewal and apoptosis that allow more efficient expansion of SKPs.

The Protective Effect of Indole-3-Acetic Acid (IAA) on H2O2-Damaged Human Dental Pulp Stem Cells Is Mediated by the AKT Pathway and Involves Increased Expression of the Transcription Factor Nuclear Factor-Erythroid 2-Related Factor 2 (Nrf2) and Its Downstream Target Heme Oxygenase 1 (HO-1).

Indole-3-acetic acid (IAA) is the most common plant hormone of the auxin class and is known to have many effects including cell proliferation enhancement and antioxidant property. However, no study has revealed its defensive effects against oxidative toxicity in human dental pulp stem cells (hDPSCs). In this study, we investigated the effects of IAA on hydrogen peroxide- (H2O2-) induced oxidative toxicity in hDPSCs. H2O2-induced cytotoxicity was attenuated after IAA treatment. Cell cycle analysis using FACS showed that the damaged cell cycle and increased number of apoptotic cells by H2O2 treatment were recovered after the treatment of IAA. The H2O2-mediated increased expression of the proapoptotic genes, BAX and p53, was attenuated by IAA treatment, while IAA treatment increased antiapoptotic genes, BCL-2 and ATF5 expression. The increases of cleaved caspase-3 and ROS by H2O2 were also decreased after treatment of IAA. To further investigate the mechanism of IAA, Nrf2-related antioxidant pathway was examined and the results showed that the level of Nrf2 and HO-1 expressions, stimulated by H2O2, decreased after treatment of IAA. Moreover, IAA treatment protected hDPSCs against H2O2-induced oxidative stress via increased expression of Nrf2 and HO-1, mediated by the AKT pathway.

Effect of Inner Taper Angle of Acetabular Metal Shell on the Malseating and Dissociation Force of Ceramic Liner.

BACKGROUND: Malseating of ceramic liner appeared as a matter of concern in multibearing metal, although multibearing cup can be coupled with hard liners as well as polyethylene liner. In this metal shell, the inner taper angle should be 10° for the modularity, while standard metal shells for ceramic liner have an inner taper angle of 18°. However, there has been no study in the effect of taper angle to the risk of malseating. We evaluated whether the taper angle of metal shell might affect the malseating of ceramic liner, and dissociation force of ceramic liner from metal shell. METHODS: Three surgeons manually inserted ceramic liners into two designs of metal shell with different tapers angles (10° and 18°). We compared malseating rates of ceramic liners and push-out strengths, which means dissociation force of the ceramic liner from the metal shell, between these two metal shell designs. RESULTS: The malseating rates in 10° metal shell were higher than those in 18° metal shell (23.3% vs 0%, P < .05). The mean dissociation force (1148.8 ± 46.7 N) in 10° taper cup was higher than that (389.7 ± 108.3 N) in 18° taper cup (P < .01). CONCLUSION: Our results suggest that surgeon should be cautious about malseating of ceramic liner when using multi-bearing metal shell with inner taper angle of 10°. CLINICAL RELEVANCE: When surgeon use multi-bearing metal shell with inner taper angle of 10°, our results suggest that surgeon should be cautious about malseating of ceramic liner.

Facile fabrication of properties-controllable graphene sheet.

Graphene has been received a considerable amount of attention as a transparent conducting electrode (TCE) which may be able to replace indium tin oxide (ITO) to overcome the significant weakness of the poor flexibility of ITO. Given that graphene is the thinnest 2-dimensional (2D) material known, it shows extremely high flexibility, and its lateral periodic honeycomb structure of sp(2)-bonded carbon atoms enables ~2.3% of incident light absorption per layer. However, there is a trade-off between the electrical resistance and the optical transmittance, and the fixed absorption rate in graphene limits is use when fabricating devices. Therefore, a more efficient method which continuously controls the optical and electrical properties of graphene is needed. Here, we introduce a method which controls the optical transmittance and the electrical resistance of graphene through various thicknesses of the top Cu layers with a Cu/Ni metal catalyst structure used to fabricate a planar mesh pattern of single and multi-layer graphene. We exhibit a continuous transmittance change from 85% (MLG) to 97.6% (SLG) at an incident light wavelength of 550 nm on graphene samples simultaneously grown in a CVD quartz tube. We also investigate the relationships between the sheet resistances.

Optimization for the removal of orthophosphate from aqueous solution by chemical precipitation using ferrous chloride.

The precipitation reaction between the orthophosphate and Fe2+ ions was studied to describe the optimum condition for the removal of orthophosphate from the aqueous solution. The effects of pH, Fe:P molar ratio, and alkalinity were evaluated for the initial orthophosphate concentrations in the range from 1.55 to 31.00 mg/L - PO4(3-) -P. The optimum pH was found to be 8.0 in all orthophosphate concentration ranges. When the stoichiometric moles of Fe2+ were added, the removal efficiencies were significantly less than the theoretical values. It is likely that the precipitation of Fe(OH)2(s) is partially formed. For the initial orthophosphate concentration of 3.10 mg/L PO4(3-) -P or greater, the removal efficiencies with the Fe:P molar ratio of 3.0:1.0 approached to the theoretical values, yielding greater than 98.5%. If the molar ratio of Fe:P was great enough, the precipitation reaction was completed within 1 h. As the alkalinity increases, the experimental removal efficiencies were significantly greater than the theoretical values. This is because the formation of vivianite is favoured over FeCO3(s). Finally, it was demonstrated that the orthophosphate (1.40-6.80 mg/L PO4(3-) -P) in the secondary effluents from wastewater treatment plants was effectively removed by dosing sufficient amount of Fe2+ ions.

Flexible and transparent gas molecule sensor integrated with sensing and heating graphene layers.

Graphene leading to high surface-to-volume ratio and outstanding conductivity is applied for gas molecule sensing with fully utilizing its unique transparent and flexible functionalities which cannot be expected from solid-state gas sensors. In order to attain a fast response and rapid recovering time, the flexible sensors also require integrated flexible and transparent heaters. Here, large-scale flexible and transparent gas molecule sensor devices, integrated with a graphene sensing channel and a graphene transparent heater for fast recovering operation, are demonstrated. This combined all-graphene device structure enables an overall device optical transmittance that exceeds 90% and reliable sensing performance with a bending strain of less than 1.4%. In particular, it is possible to classify the fast (≈14 s) and slow (≈95 s) response due to sp(2) -carbon bonding and disorders on graphene and the self-integrated graphene heater leads to the rapid recovery (≈11 s) of a 2 cm × 2 cm sized sensor with reproducible sensing cycles, including full recovery steps without significant signal degradation under exposure to NO2 gas.

SnO2 nanoslab as NO2 sensor: identification of the NO2 sensing mechanism on a SnO2 surface.

Among the various metal oxides, SnO2 has been most widely exploited as a semiconductor gas sensor for its excellent functionalities. Models illustrating the sensing mechanism of SnO2 have been proposed and tested to explain experimentally derived "power laws". The models, however, are often based on somewhat simplistic assumptions; for instance, the net charge transfer from an adsorbate to a sensor surface site is assumed to occur only by integer values independent of the crystallographic planes. In this work, we use layer-shaped SnO2 crystallites with one nanodimension (1ND-crystallites) as NO2 gas sensing elements under flat band conditions, and derive appropriate "power laws" by combining the dynamics of gas molecules on the sensor surface with a depletion theory of semiconductor. Our experimentally measured sensor response as a function of NO2 concentration when compared with the theoretically derived power law indicates that sensing occurs primarily through the chemisorption of single NO2 molecules at oxygen vacancy sites on the sensor surface.

Evaluation for protein binding affinity of maghemite and magnetite nanoparticles.

We investigated the protein binding affinity of magnetite (Fe3O4) and maghemite (gamma-Fe2O3) nanoparticles with against non-characterized protein from human lung cancer A549 cell line on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The binding ability of maghemite was 400 ng/mg. According to the SDS-PAGE results, the protein binding affinity of maghemite nanoparticles is stronger than magnetite nanoparticles. These data suggest that a protein can be detected with maghemite nanoparticles.

Identification of origin of unknown derivative chromosomes by array-based comparative genomic hybridization using pre- and postnatal clinical samples.

Microarray-based comparative genomic hybridization (array CGH) is a high-resolution and comprehensive method for detecting both genome-wide and chromosome-specific copy-number imbalance. We have developed an array CGH analysis system (consisting of an array CGH chip plus its exclusive analysis software) for constitutional genetic diagnosis and have evaluated the suitability of our system for molecular diagnosis using pre- and postnatal clinical samples. In a blind study, each of the 264 sample karyotypes identified by array CGH analysis was consistent with that identified by traditional karyotype analysis--with one exception, case (47, XXX)--and we were able to identify origins, such as small supernumerary marker chromosomes, which cannot be determined by conventional cytogenetics. We also acquired very accurate, fast and reliable results using a diminutive amount of clinical samples. Taken together, the array CGH platform developed in this study is a rapid, powerful and sensitive technology for pre- and postnatal diagnosis using a very small amount of clinical sample.

Fetal gender determination and BclI polymorphism using nucleated erythrocytes in maternal blood.

This study demonstrated determination of fetal gender from nucleated red blood cells (NRBCs) in maternal blood and attempted to apply prenatal diagnosis of hemophilia A using BclI DNA polymorphism. Venous blood was drawn from 20 pregnant women, and NRBCs were recovered by magnetic activated cell sorting and anti-GPA (glycophorin A) immunostaining. After microdissector isolation of the NRBCs, primer extension preamplification (PEP) and nested PCR of the amelogenin gene were performed to determine fetal gender. We also performed PEP and nested PCR of BclI polymorphism to verify the validity of prenatal diagnosis of hemophilia A. DNA amplification was achieved in 107 cells (51.9%) and fetal gender determined with 65.0% accuracy. Unfortunately, we could not verify the validity within the scope of this study. However, in a larger number of cases that are informative in BclI polymorphism, we will be able to identify patients affected by hemophilia A using fetal NRBCs in maternal blood.